Juan A. Correa

Red algal farming in Chile: a review

Abstract Production of seaweeds in Chile has fluctuated between 74,000 and 322,000 wet metric tons/year during the last 14 years, involving different species of Phaeophyta and Rhodophyta. Among Rhodophyta, the most important harvested species include the carrageenophytes Sarcothalia crispata, Mazzaella laminarioides, Gigartina skottsbergii, Chondracanthus chamissoi, and the agarophytes Gracilaria chilensis and Gelidium lingulatum. Other less important taxa are Gel. rex, M. membranacea, Ahnfeltia plicata, Ahnfeltiopsis furcellata, Porphyra columbina, Callophyllis variegata, Mastocarpus papillatus and Chondrus canaliculatus. Chilean production comes mainly from wild stocks, as at present, cultivation on a commercial scale is restricted to Gra. chilensis. Total landings of Gracilaria currently stand at 120,000 wet tons. Large-scale cultivated biomass of this species, on the other hand, has been the result of a sharp increase in the number of farms, from less than 10 in 1982 to almost 322 in 1996. A basic understanding of key biological and ecophysiological aspects, as well as the availability of propagation methods, permitted the development of large-scale Gracilaria farming operations. However, during the cultivation process, new problems arose for the farmers, such as abrupt production decline, pests and pathogens. Similar key knowledge is lacking for other Chilean Rhodophyta, which creates a bottleneck that prevents the development of seaweed farming activities other than Gracilaria. This situation prevails in spite of the growing pressure on wild stocks triggered by an increase in the demand for raw material by the industry, with the obvious danger of over-exploitation and the resulting collapse of fisheries. Taking the above into consideration, an effort has been made in recent years to provide the basic knowledge necessary for the management and cultivation of some of the most valuable seaweed resources in Chile. Thus, the main objective of this contribution is to summarize the present situation of red seaweed cultivation in the country. We will address this issue by reviewing the landing statistics of these resources, followed by a summary of recent information that favours cultivation. These include propagation methods, culture conditions and techniques, product quality, pest management, strain selection and the diversification of seaweeds currently exploited in Chile.

Sulfated Oligosaccharides Mediate the Interaction between a Marine Red Alga and Its Green Algal Pathogenic Endophyte

The endophytic green alga Acrochaete operculata completely colonizes the sporophytes of the red alga Chondrus crispus; however, it does not penetrate beyond the outer cell layers of the gametophytes. Given that the life cycle phases of C. crispus differ in the sulfation pattern of their extracellular matrix carrageenans, we investigated whether carrageenan fragments could modulate parasite virulence. λ-Carrageenan oligosaccharides induced release of H2O2, stimulated protein synthesis, increased carrageenolytic activity, and induced specific polypeptides in the pathogen, resulting in a marked increase in pathogenicity. In contrast, κ-carrageenan oligosaccharides did not induce a marked release of H2O2 from A. operculata but hindered amino acid uptake and enhanced their recognition by the host, resulting in a reduced virulence. Moreover, C. crispus life cycle phases were shown to behave differently in their response to challenge with cell-free extracts of A. operculata. Gametophytes exhibited a large burst of H2O2, whereas only low levels were released from the sporophytes.

Phylogeographic analyses of the 30°S south-east Pacific biogeographic transition zone establish the occurrence of a sharp genetic discontinuity in the kelp Lessonia nigrescens: Vicariance or parapatry?

Phylogeographic studies are lacking in the Southern Hemisphere, and in particular in the south-eastern Pacific. To infer the possible scenario for the debated biogeographic transition zone located at 30-33 degrees S along the Chilean coast, we investigated whether there is a concordance between the phylogeographic pattern and the biogeographic transition in the intertidal kelp Lessonia nigrescens whose distribution is continuous across this transition zone. Using a combination of four makers located in the three genomic compartments (chloroplast, mitochondria and nucleus), we showed the presence of two main divergent lineages, possibly cryptic species. There was an exact match of the phylogeographic break with the 30 degrees S biogeographic transition zone, suggesting a common origin. The combined information given by the multilocus approach and by the population analysis suggested the occurrence of a budding speciation, with a northward range expansion.

Differential responses to copper-induced oxidative stress in the marine macroalgae Lessonia nigrescens and Scytosiphon lomentaria (Phaeophyceae)

In order to help explain the absence of the brown kelp Lessonia nigrescens from a coastal environment chronically enriched with copper, we characterized the biochemical responses induced by copper stress in this kelp and compared them with those displayed by the copper tolerant brown alga Scytosiphon lomentaria. These algae were cultivated with increasing concentrations of copper (20, 40 and 100microgL(-1)) for 96h and the temporal production of hydrogen peroxide, superoxide anions and lipoperoxides as well as the activities of antioxidant enzymes catalase (CAT), glutathione peroxidase (GP), ascorbate peroxidase (AP), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) and the activity of the defense enzyme lipoxygenase (LOX) were determined. In L. nigrescens and S. lomentaria, a single peak of hydrogen peroxide was detected, with similar maxima after 3h of copper exposure, although in L. nigrescens buffering took longer. Superoxide anions, on the other hand, were only detected in L. nigrescens. The production of lipoperoxides in L. nigrescens increased steadily at higher copper levels, in a pattern clearly different to their rapid stabilization in S. lomentaria. We suggest that the accumulation of lipoperoxides might be related to LOX, whose activity also increases with exposure time. Furthermore, activities of the antioxidant enzymes CAT, GP, AP and DHAR were lower in L. nigrescens than in S. lomentaria, and GP and DHAR were completely inhibited at higher copper concentrations. Since these enzymes also detoxify fatty acid hydroperoxides, their inhibition, together with the activation of LOX, may explain the persistent and copper-dependent levels of lipoperoxides in L. nigrescens. Based on terrestrial plant models demonstrating toxic effects of lipoperoxides, and on our results on organellar ultrastructural changes, we suggest that copper toxicity induced an uncontrolled lipoperoxide accumulation which may lead to cell damage and dysfunction in L. nigrescens, explaining at least partially, the absence of this kelp in a copper-enriched coastal environment.

Copper stress induces biosynthesis of octadecanoid and eicosanoid oxygenated derivatives in the brown algal kelp Laminaria digitata.

To better understand the toxicity and the orchestration of antioxidant defenses of marine brown algae in response to copper-induced stress, lipid peroxidation processes were investigated in the brown alga Laminaria digitata. The expression of genes involved in cell protection and anti-oxidant responses were monitored by semi-quantitative reverse transcriptase polymerase chain reaction and the lipid peroxidation products were further characterized by profiling oxylipin signatures using high-pressure liquid chromatography-mass spectrometry. Exposure to copper excess triggers lipoperoxide accumulation and upregulates the expression of stress related genes. It also increases the release of free polyunsaturated fatty acids, leading to an oxidative cascade through at least two distinct mechanisms. Incubations in presence of inhibitors of lipoxygenases and cycloxygenases showed that in addition to the reactive oxygen species-mediated processes, copper stress induces the synthesis of oxylipins through enzymatic mechanisms. Among complex oxylipins, cyclopentenones from C18 and C20 fatty acids such as 12-oxo-PDA and prostaglandins were detected for the first time in brown algae, as well as unique compounds such as the 18-hydroxy-17-oxo-eicosatetraenoic acid. These results suggest that lipid peroxidation participates in the toxic effects of copper and that lipid peroxidation derivatives may regulate protective mechanisms by employing plant-like octadecanoid signals but also eicosanoid oxylipins which are absent in vascular plants.

CONVERGENT BIOLOGICAL PROCESSES IN COALESCING RHODOPHYTA

Sporeling coalescence in Gracilaria chilensis Bird, McLachlan et Oliveira produces genetically polymorphic, chimeric individuals. If this is common in red algae, it may have significant biological consequences. In this study, we evaluate the hypotheses that coalescence is widespread among the Rhodophyta and that specific and convergent morphological and ecological responses characterize this as a unique growth style among marine algae. A literature survey on coalescence was undertaken to assess the distribution of this condition in the Florideophycidae. Sixty‐two (54.9%) of 113 species considered germinated to form a disk. Subsequent development in 37 of these species showed crust formation and coalescence during development with other crusts in 31 species (84%). Coalescing red algae were members of the orders Ahnfeltiales, Corallinales, Gigartinales, Gracilariales, Halymeniales, Palmariales, and Rhodymeniales. Ultrastructural studies in species of Ahnfeltiopsis, Chondrus, Gracilaria, Mazzaella, and Sarcothalia suggested a common pattern of early development. Newly released, naked spores may fuse into a single cell, as they do in Chondrus canaliculatus, or they may develop individual cell walls that later are surrounded by a thickened common wall. Ultrastructural studies demonstrated two kinds of immediate development after the first mitotic division: direct development by symmetric divisions resulting in discoid sporelings or an indirect asymmetric arrangement of divisions before a diskoid sporeling was formed. Germination in coalescing species is a linear function of the initial spore density, whereas in noncoalescing species maximum germination occurs at intermediate densities. In the field, coalescing species may recruit either from solitary or aggregated spores. However, survival is significantly higher for plantlets grown from a larger number of coalescing spores. Total number of erect axes formed by the coalesced mass is a logarithmic function of the initial number of spores. Thus, germlings grown from a larger number of coalescing spores exhibited a larger photosynthetic canopy than do plantlets grown from a few spores. Juveniles and mature clumps grown from a coalescing mass may exhibit size inequalities among erect axes, with the larger axes located toward the center of the clump. These larger axes mature first or, in some cases, are the only to produce spores. The widespread occurrence of coalescence inroughly half the number of orders of the Florideophycidae, the similarity of the coalescence process, and the finding of various adaptive traits associated with coalescence characterizes this as a unique growth style, splitting the diversity of species now included in the Florideophycidae into two major groups: coalescing and noncoalescing Rhodophyta.

Copper, copper mine tailings and their effect on marine algae in Northern Chile

Results are presented of a long-term research programme on the effect of copper contamination on biota in Chilean coastal waters. In spite of the magnitude of the copper mining tailings that affected Caleta Palito and surroundings in northern Chile, the effects on the intertidal assemblages remain restricted to a small geographic area. Even within the affected area, the effects are not homogeneous and there is evidence of active recovery in biological diversity in recent few years. Experimental evidence suggests that the current low algal diversity and abundance is strongly influenced by herbivory, although chronic effects of the discharges cannot be ruled out. Cellular changes in Enteromorpha compressa from the impacted area were characterised by abnormal granules in the cytoplasm, though these granules did not contain detectable levels of copper or other heavy metals.

Copper stress proteomics highlights local adaptation of two strains of the model brown alga Ectocarpus siliculosus.

Ectocarpus siliculosus is a cosmopolitan brown alga with capacity to thrive in copper enriched environments. Analysis of copper toxicity was conducted in two strains of E. siliculosus isolated from (i) an uncontaminated coast in southern Peru (Es32) and (ii) a copper polluted rocky beach in northern Chile (Es524). Es32 was more sensitive than Es524, with toxicity detected at 50 μg/L Cu, whereas Es524 displayed negative effects only when exposed to 250 μg/L Cu. Differential soluble proteome profiling for each strain exposed to sub‐lethal copper levels allowed to identify the induction of proteins related to processes such as energy production, glutathione metabolism as well as accumulation of HSPs. In addition, the inter‐strain comparison of stress‐related proteomes led to identify features related to copper tolerance in Es524, such as striking expression of a PSII Mn‐stabilizing protein and a Fucoxanthine chlorophyll a–c binding protein. Es524 also expressed specific stress‐related enzymes such as RNA helicases from the DEAD box families and a vanadium‐dependent bromoperoxidase. These observations were supported by RT‐qPCR for some of the identified genes and an enzyme activity assay for vanadium‐dependent bromoperoxidase. Therefore, the occurrence of two different phenotypes within two distinct E. siliculosus strains studied at the physiological and proteomic levels strongly suggest that persistent copper stress may represent a selective force leading to the development of strains genetically adapted to copper contaminated sites.

Antioxidant responses in Scytosiphon lomentaria (Phaeophyceae) inhabiting copper-enriched coastal environments

Scytosiphon lomentaria (Lingb.) Link. (Phaeophyceae) is one of the two dominant seaweeds in a coastal area of northern Chile affected by copper mine wastes, where the concentration of copper in water and algal tissues remains higher than in nonimpacted sites. Copper‐loaded plants develop oxidative stress, as demonstrated by the increased levels of reactive oxygen species and lipoperoxides. This stress was associated with 1) an enhanced activity of the antioxidant enzymes catalase, glutathione peroxidase, ascorbate peroxidase, monodehydroascorbate reductase, and dehydroascorbate reductase and 2) an inhibition of the glutathione reductase activity. Furthermore, stressed plants showed a decrease in glutathione and phenolic compounds levels and an increase in total ascorbate. Reciprocal transplants revealed that plants rapidly adjusted their antioxidant system in response to the conditions of the receiving site. In individuals transplanted from the copper‐enriched environment to the control site, normal levels of lipoperoxides and antioxidant compounds were restored in 48 h and antioxidant enzymes recovered their basal activities in 96 h. Individuals transplanted from the control site to the copper‐enriched area adjusted their antioxidant compounds and antioxidant enzymes within 48 h and 96 h, respectively, and reached the functional status of the local plants. We conclude that S. lomentaria inhabiting the copper‐enriched area buffered oxidative stress by a simultaneous involvement of antioxidant enzymes and water‐soluble antioxidant compounds. These antioxidant responses were rapid and reversible, suggesting that copper resistance in S. lomentaria is a constitutive trait and that copper enrichment of the area did not result in a locally adapted copper‐tolerant ecotype.

Tolerance to oxidative stress induced by desiccation in Porphyra columbina (Bangiales, Rhodophyta)

Unravelling the mechanisms underlying desiccation tolerance is crucial in order to understand the position of algal species in the intertidal zone. The alga Porphyra columbina lives in the uppermost part of the rocky intertidal zones around the world and was selected as a model for this study. Naturally desiccated plants were collected during low tide and studied for morphological changes, oxidative burst induction, biomolecule oxidation, antioxidant responses, and photosynthetic status. Naturally hydrated plants collected during high tides were used for comparative purposes. In addition, changes induced by desiccation were assessed in vitro and the capacity to recover from desiccation was determined by rehydrating the fronds in seawater. The global results show that desiccation induces morphological and cellular alterations accompanied by a loss of ∼96% of the water content. Overproduction of reactive oxygen species (ROS) was induced by desiccation and two peaks of H2O2 were detected at 1 and 3 h of desiccation. However, during in vitro rehydration post-desiccation, the ROS quickly returned to the basal levels. At the biomolecular level, only a low production of oxidized proteins was recorded during desiccation, whereas the activity of diverse antioxidant enzymes increased. However, this activity diminished to near basal levels during rehydration. The photosynthetic efficiency (Fv/Fm) during desiccation declined by 94–96% of the values recorded in hydrated plants. This reduction was generated by the low levels of trapped energy flux per cross-section (TRo/CS), electron transport flux per CS (ETo/CS), and density of reaction centres (RC/SCo) as well as the chlorophyll content. The inverse pattern was observed for the levels of phycocyanin and phycoerythrin content. Fv/Fm and the photosynthetic indicators were restored to normal levels after only 5 min of rehydration. The results indicate that desiccation in P. columbina causes overproduction of ROS that is efficiently attenuated. The morphological and photosynthetic changes could be operating as tolerance mechanisms due to the fact that these responses principally prevent biomolecular alteration and cellular collapse. Thus, the activation of different physiological mechanisms helps to explain the high tolerance to desiccation of P. columbina and, at least in part, the position of this species at the highest level in the intertidal zone.